The identified key residues might either directly contact Gn/Gc or might indirectly contribute to Gn/Gc binding through intramolecular interactions that maintain the local geometry of EC1, as is likely the case for more buried residues (e.g., L152). 34, 36983707 (2006). 1a) and mapped this human-murine difference in susceptibility to a sequence variation at a single residue in EC1, residue 83, that modulates Gn/Gc-PCDH1 engagement (Fig. https://doi.org/10.1038/s41467-023-40126-y, DOI: https://doi.org/10.1038/s41467-023-40126-y. How is hantavirus spread? " Hantaviruses: A Global Disease Problem ." Emerging Infectious Diseases, 3.2 (June 1997). CAS Soluble PCDH1 variants cloned into pcDNA3.1 (see above) were expressed in 293F cells in shaker flasks by transient transfection with linear polyethyleneimine (Polysciences) and purified by nickel-chelation chromatography. [35] Schilling and colleagues have clearly shown the limits of serotyping by nonfocus reduction neutralization test methods. is a member of the scientific advisory board of Integrum Scientific LLC and has consulted for Axon Advisors, LLC. 2c, d) was concordant with the essentially complete loss in its binding to sEC1-2(F83L) observed in the direct binding ELISA (Fig. Virol. Please see our. MeansSD: n=9 wells of infected cells examined over three experiments. After thoroughly washing with PBST, western blots were imaged on a LI-COR Odyssey Fc Imager (LI-COR Biosciences, LICOR Image Studio software, V.1.0.19) and the uncropped images are included in the Source Data file. lower right]. Replication-competent, recombinant vesicular stomatitis Indiana viruses (rVSVs) expressing an eGFP reporter and bearing VSV G or hantavirus Gn/Gc of either ANDV (NP_604472.1), Sin Nombre virus (SNV) (NP_941974.1), or Hantaan virus (HTNV) (NP_941978.1) with the modifications described in Slough et al.42, were generated using a plasmid-based rescue system in 293T cells and propagated on Vero cells as described previously43,44. Averages SEM: n=6 wells of each dilution examined over three independent experiments, except for: sEC1-2(D102A, K104A, K104E, T105A, V144A, Q145A, D102R, D142A, G75R, Y81A, P84A, I140A) have n=8 wells examined over four independent experiments [sEC1-2(Y81) has n=7 wells for one dilution]. Serum samples were serially diluted in 5% milk protein in PBS/0.02% Tween 20 and added to antigen-coated plates (2h at RT). Next, we evaluated the roles of two individual residuesF83, which influences cellular susceptibility to viral entry (Fig. Infectivities (c, f) were compared by one-way ANOVA with Dunnetts correction for multiple comparisons; ns>0.05, **P<0.01. If material is not included in the articles Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. Contact the LMR via the paging officer at 206-598-6190. volume14, Articlenumber:4454 (2023) Previous work has shown that the PCDH1 ectodomain can form head-to-tail homodimers, driven largely by intermolecular interactions between the EC1 and EC4 domains17 (Fig. Here, we found that hantaviruses recognize a surface in PCDH1 EC1 that is distinct from the EC1-EC4 adhesive interface (Fig. The hantavirus surface glycoprotein lattice and its fusion control mechanism. Commenting is limited to medical professionals. Linking to a non-federal website does not constitute an endorsement by CDC or any of its employees of the sponsors or the information and products presented on the website. Please enter a Recipient Address and/or check the Send me a copy checkbox. Hantaviruses, from the Bunyaviridae family, are a group of viruses that are normally carried by rodents, such as rats, mice and voles. Hooper, J. W., Larsen, T., Custer, D. M. & Schmaljohn, C. S. A lethal disease model for hantavirus pulmonary syndrome. As reported previously, PCDH1s EC1 is necessary and sufficient for binding by New World hantavirus Gn/Gc, and PCDH1 loss greatly reduces the lethality of ANDV infection in a Syrian hamster model of HCPS6. The results obtained with this new technique were compared with those obtained using the strip immunoblot assay from US CDC, demonstrating a high sensitivity. J. Trop. or PCDH1(F83A/D85R)) Syrian golden hamsters (Utah State University), 5-12 weeks old, male and female, were exposed to 2000 PFU of ANDV strain Chile-9717869 diluted in PBS, via the intranasal route and monitored for up to 35 days post-exposure. Infected deer mice live throughout the state and people are at risk for HPS in any part of Washington. This video will assist individuals whose work may bring them into contact with rodents. 2). Source data are provided as a Source Data file. Rankings of the residues in the PCDH1 ectodomain for each of the five complimentary algorithms can be found in Supplementary Data2. This training video was developed in partnership with Navajo Nation, Indian Health Services, and the Arizona and New Mexico Departments of Health. 2e): both sEC1-2(WT) and sEC1-2(F83L) could block rVSV-ANDV-Gn/Gc infection, but the latter was less active (Fig. J. Mol. Source data are provided as a Source Data file. Cite this article. Retroviruses packaging the transgenes were produced by transfecting 293T cells43, and target cells were directly exposed to sterile-filtered, retrovirus-laden supernatants in the presence of polybrene (6g/mL). A new rapid diagnostic method for hantavirus has been developed based on a highly dispersed immunoelectrode prototype amperometric immunosensor. A peer review file is available. Prevention How to prevent hantavirus infections. Sci. A representative blot from a single experiment of two independent experiments is shown. Individuals with symptoms of hantavirus infection and a history of exposure to a rodent or its droppings, urine, or nest, should be tested for confirmation. 3b, c and 5c, d). 3) onto sEC1-2coated ELISA plates (Fig. Spread. CRL-3216), grivet kidney Vero cells (ATCC, catalog No. By contrast, and consistent with the higher affinity of the ANDV Gn/Gc:PCDH1 interaction, a combination of both mutations was necessary to reduce ANDV Gn/Gc-mediated infection (Fig. Seroconversion, indicative of an ANDV-specific IgG response, was observed in all surviving hamsters, indicating they had all been exposed to the virus (Supplementary Fig. We chose to evaluate ANDV and not SNV in these screens because the weaker binding affinity of the latter Gn/Gc for PCDH1 necessitated prohibitive quantities of viral material for binding assays and was expected to reduce our capacity to detect nuanced reductions in binding by mutant sEC1-2 proteins. For detection of hantavirus infections in rodents and insectivores, serology followed by immunoblotting of, for example, lung tissue, and RT-PCR detection of viral RNA may be used, and if of interest followed by sequencing and virus isolation. Figure(a) includes an image from Flaticon.com. 7a), responsible for driving the EC1:EC4 interaction17, on ANDV Gn/Gc binding. Later symptoms include trouble breathing and a rapid heartbeat. Infectivities were compared by one-way ANOVA with Dunnetts test for multiple comparisons. Contact the LMR via the paging officer at 206-598-6190. Treatment focuses on helping you breathe and reducing your symptoms. HPS can be fatal if not diagnosed and treated quickly. b Hierarchical clustering of WT and mutant sEC1-2 generated from sigmoidal curves of the infection-inhibition assay in (a). Alex Martins Machado; Glauciane Garcia de Figueiredo; Gilberto Sabino dos Santos Jr; Luiz Tadeu Moraes Figueiredo. Safronetz, D., Ebihara, H., Feldmann, H. & Hooper, J. W. The Syrian hamster model of hantavirus pulmonary syndrome. Sera are initially screened for IgG and IgM antibodies recognizing the nucleocapsid protein common to all hantaviruses. d EC1 crystal structure in the open conformation displaying mutated residues representing three degrees of binding strength to ANDV Gn/Gc and inhibition of rVSV-ANDV-Gn/Gc infection. Andes virus (ANDV) and Sin Nombre virus (SNV) are the etiologic agents of severe hantavirus cardiopulmonary syndrome (HCPS) in the Americas for which no FDA-approved countermeasures are available. Herein, we link the inter-species sequence variations in PCDH1 and mammalian host-specific receptor usage by hantaviruses to pinpoint sequences that could influence susceptibility to hantavirus infection. POC-PUUMALA is a new immunochromatographic assay for hantavirus. For PCDH1 surface expression, MLMECs and human U2OS cells expressing variant PCDH1 were seeded on fibronectin-coated glass coverslips 24h pre-immunostaining. CCL-81), and grivet kidney Vero E6 cells (ATCC, catalog No. Groups of wild-type Syrian golden hamsters (Mesocricetus auratus) (Envigo) and PCDH1-KI (PCDH1(10a.a.) Lassa virus entry requires a trigger-induced receptor switch. The two most divergent loop conformations were selected as representative loop conformations to perform structure-based interfacial residue predictions on. Biallelic PCDH1(F83A/D85R) animals expressed levels of PCDH1 in the lung similar to their WT counterparts (Fig. Virology 289, 614 (2001). c Area under the curve (AUC) for the binding activity of WT and mutant sEC1-2 to rVSV-ANDV-Gn/Gc, as determined by competition ELISA (see Fig. Health professionals in Canada are advised to be vigilant. The Centers for Disease Control and Prevention (CDC) cannot attest to the accuracy of a non-federal website. 7a). Internet Explorer). To test ARAV rN as an antigen for antibody detection, sera from 30 patients from Argentina, previously known as seropositive for hantavirus, were tested and were all found to be positive for IgG and IgM by ELISA, using either ARAV or Andes vrus (ANDV) rN antigens. Although the EC4 and Gn/Gc contact sites in EC1 do not appear to overlap, we considered the possibility that EC1 dimerization may nevertheless impact Gn/Gc recognition. Source data are provided as a Source Data file. 7c). An ISH probe targeting ANDV S segment (GenBank accession number: NC_003466.1) was designed and synthesized by Advanced Cell Diagnostics (#900241, Advanced Cell Diagnostics). Med. MeansSEM: n=7 wells examined over three independent experiments (ANDV), n=4 wells examined over two independent experiments (SNV). Scale bars represent 100m. Hantaviruses are a group of viruses that may be carried by some rodents. Animals were observed daily for clinical signs of disease, morbidity, and mortality. PCDH1 interaction and homodimerization in trans is proposed to regulate cell adhesion between neighboring cells, suggesting that both monomers and dimers co-exist at the cell surface17. The Gn/Gc binding site relative to the EC1:EC4 adhesive interface (dark blue) is indicated. Specifically, we observed that murine endothelial cells are refractory to SNV Gn/Gc-dependent entry (Fig. Following a final wash, 2,2-Azinobis [3-ethylbenzothiazoline-6-sulfonic acid]-diammonium salt (ABTS) substrate (Kirkegaard and Perry Laboratories, Inc.) was added and absorbance values were read at 405nm using a Spectramax plate reader (Molecular Devices, LLC). Cifuentes-Muoz, N., Salazar-Quiroz, N. & Tischler, N. D. Hantavirus Gn and Gc envelope glycoproteins: key structural units for virus cell entry and virus assembly. The Hantavirus genome can be detected by reverse transcription (RT)-PCR in clinical samples. 25, 159165 (2016). PCDH1 directly engages the tetrameric viral Gn/Gc glycoprotein complex6,7, which mediates cell entry8. Infection with any hantavirus can produce hantavirus disease in people. [2224] The N antigenic sites, conserved among those of the Hantavirus genus, are located in the aminoproximal region of the protein, based on studies with PUUV, Hantaan virus (HTNV) and Sin Nombre virus (SNV). 1cd), indicating that a molecular incompatibility between SNV Gn/Gc and murine PCDH1 is at least partially responsible for the entry block in murine endothelial cells. Further information on research design is available in theNature Portfolio Reporting Summary linked to this article. Instead, the differences in binding affinity between ANDV and SNV Gn/Gc (and by extension, the glycoproteins from other New World hantaviruses) likely arise from sequence variations in Gn/Gcs yet-unmapped PCDH1-binding site (also see below). All Hanta IgM positive samples from US residents will be sent to a Public Health Laboratory for SNV-specific IgM.